College of Animal Science and Technology, Anhui Agricultural University, Hefei, China.
Fish Shellfish Immunol. 2013 Jan;34(1):291-5. doi: 10.1016/j.fsi.2012.11.015. Epub 2012 Nov 23.
Vibrio mimicus (V. mimicus) is the causative agent of ascites disease in aquatic animals. Outer membrane protein U (OmpU) is an important antigen of V. mimicus, but its protective epitopes are still unclear. A random 12-mer phage-displayed peptide library was used to screen and identify immunodominant mimotopes of the OmpU protein in V. mimicus by panning against purified OmpU-specific polyclonal antibody. Then the immunogenicity and immunoprotection in fish of these mimotopes was evaluated. Nine positive phage clones presented seven different 12- peptide sequences and more than 50% of them carried a consensus core motif of DSSK-P. These positive clones reacted with the target antibody and this interaction could be blocked, in a dose-dependent manner, by OmpU protein. Intraperitoneal injection of seven positive phage clones into fish induced a specific antibody response to OmpU protein. The fish immunized respectively with the positive phage clones C17, C24, C60 and C66 obtained 100% immunoprotective effect against experimental V. mimicus challenge. Taken together, these mimotopes presented by clone C17, C24, C60 and C66 were immunodominant mimotopes of the OmpU protein and exhibited a more appropriate candidate as epitope-based vaccine against V. mimicus infection in aquatic animals.
威氏弧菌(Vibrio mimicus)是水生动物腹水病的病原体。外膜蛋白 U(OmpU)是威氏弧菌的重要抗原,但它的保护性表位仍不清楚。本研究通过用纯化的 OmpU 特异性多克隆抗体淘选,从随机 12 肽噬菌体展示肽库中筛选和鉴定威氏弧菌 OmpU 蛋白的免疫显性模拟肽。然后评估这些模拟肽在鱼类中的免疫原性和免疫保护作用。通过对纯化的 OmpU 特异性多克隆抗体进行淘选,从随机 12 肽噬菌体展示肽库中筛选和鉴定了威氏弧菌 OmpU 蛋白的免疫显性模拟肽。然后评估了这些模拟肽在鱼类中的免疫原性和免疫保护作用。9 个阳性噬菌体克隆呈现出 7 种不同的 12 肽序列,其中 50%以上的序列含有 DSSK-P 的保守核心基序。这些阳性克隆与靶抗体发生反应,这种相互作用可以被 OmpU 蛋白以剂量依赖的方式阻断。将 7 个阳性噬菌体克隆腹腔注射到鱼体内,可诱导针对 OmpU 蛋白的特异性抗体反应。分别用阳性噬菌体克隆 C17、C24、C60 和 C66 免疫的鱼对实验性威氏弧菌攻毒获得了 100%的免疫保护作用。综上所述,这些由克隆 C17、C24、C60 和 C66 呈现的模拟肽是 OmpU 蛋白的免疫显性模拟肽,是一种更合适的候选物,可作为针对水生动物威氏弧菌感染的基于表位的疫苗。